(A) Shown are the number of human H3N2 influenza NA sequences in the GISAID EpiFlu database with the given identity at site 151 for each year from 2000 to 2014. Since 2007, ambiguous amino-acid …
This 7-zip archive contains the source code used for Figure 1 (the analysis of mutation frequencies at site 151 in naturally occurring sequences).
(A) Pure and mixed populations were generated by reverse genetics. Cells were transfected with a Hanoi/2007 NA plasmid encoding D151, G151, or an equal mix of the two, along with isogenic plasmids …
The data here differ from Figure 2A in that the virus populations were generated by reverse genetics using the unmodified A/WSN/33 PB1 gene rather than the PB1 segment modified to package GFP. …
Presumably, this improvement occurs because oseltamivir blocks the binding of G151 NA to receptor, allowing newly formed virions to be released more efficiently. Oseltamivir also slightly increases …
Shown are the allele frequencies at NA site 151 over five tissue-culture passages of initially pure D151 viruses, pure G151 viruses, or an equal mix of the two. Each passage was seeded at a total …
This 7-zip archive contains the data and source code used for Figure 3 (the analysis of mutation frequencies at site 151 after serial passage in the lab).
The code and all the FASTQ files are also available at http://dx.doi.org/10.5061/dryad.s3rs0.
Shown are the titers after reverse genetics with the indicated variant of the Hanoi/2007 NA. The experiments here parallel those in Figure 2A. Black lines indicate the mean and standard error of the …
Cells were infected at an MOI of (A) 0.02 or (B) 0.5 with pure D151 virus, pure G151 virus, or an equal mix of the two. The total MOI of infecting virus was the same across the mixed and pure …
Cooperation occurs between the D151 and G151 NA variants in viruses with HA from the Hanoi/2007 strain, but not in viruses with HA from the Wisconsin/2005 strain. Shown are the titers after reverse …
Shown are the titers after reverse genetics with the indicated Hanoi/2007 NAs in combination with an engineered binding-deficient H3 HA with multiple mutations to the receptor-binding pocket (Hooper …
Prior reports of variation at neuraminidase site 151 when H3N2 clinical specimens are passaged in cell culture.
Reference | Summary |
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(McKimm-Breschkin et al., 2003) | Sanger sequencing of 38 oseltamivir- and zanamivir-resistant MDCK-passaged clinical isolates found that 7 had G, N, E, or V at site 151. |
(Lin et al., 2010) | Sanger sequencing of 18 isolates after passage in MDCK or MDCK-SIAT1 cells found 4 isolates as D+G, 3 as D+N, and 2 as D+A. Pyrosequencing detected low frequencies of G151 and N151 in some clinical samples. |
(Tamura et al., 2013) | Pyrosequencing of 150 isolates after 1-4 passages in MDCK cells found that 85% developed mixed populations at site 151; 29% did so after a single passage. Mixed populations consisted of D+N, D+G, D+G+N, and D+G+A genotypes. T148I/K/P mutations were also observed in 23% of isolates. |
(Lee et al., 2013) | 77 clinical specimens were Sanger-sequenced before and after a single passage in MDCK cells. 18 acquired a mutation at NA site 151: 10 were D+N, 7 were D+G and one fixed D151N at the limit of detection. No mutations were detected in the unpassaged specimens. |
(Chambers et al., 2014) | 9 A/Victoria/361/11-like clinical specimens were passaged twice in MDCK cells and Sanger-sequenced before and after expansion. 4 isolates developed NA-dependent cell binding; 3 had D151G, the other D151N. |
(Mishin et al., 2014) | Pyrosequencing of 150 MDCK-grown isolates found that 42 were D+G, 34 were D+N, and 57 were D+G+N. Pyrosequencing of 50 matched clinical specimens detected no variation at site 151. |
(Mohr et al., 2015) | 16 pairs of isolates cultured in parallel in MDCK cells and in eggs were sequenced using Ion Torrent. 5 MDCK isolates were D+N, 4 were D+G, and 2 were D+N+G. No egg-passaged isolates had mutations at site 151. T148I/K mutations were observed in 7 MDCK isolates. |
This text file contains the coding sequences for the HA and NA genes used in this study.